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目的研究疲劳模型大鼠学习记忆及海马N-甲基-D-天冬氨酸(N-methyl D-aspartate,NMDA)受体NR2A、NR2B亚基及EphB2受体表达的变化,并观察四逆散、生慧汤、四慧合剂对其影响。方法采用多平台法,进行连续睡眠剥夺168h复制中枢疲劳模型,将实验动物随机分为正常对照组、模型组、四逆散组、生慧汤组、四慧合剂组,每组10只;分别在睡眠剥夺6、30、54、78、102、126、150h时间点灌胃给药及蒸馏水。应用Y迷宫观察大鼠学习记忆能力的变化,并使用实时荧光定量PCR(Real-time PCR)技术定量分析大鼠海马NR2A、NR2B亚基及EphB2受体mRNA表达的变化。结果与正常对照组比较,模型组大鼠Y迷宫正确百分率明显下降(P<0.05),海马NR2B和EphB2的mRNA表达明显减少(P<0.01),NR2A表达无明显变化。与模型组比较,四慧合剂组能明显提高大鼠迷宫成绩和海马NR2A、EphB2的mRNA表达(P<0.01),而四逆散组、生慧汤组差异无统计学意义。与四逆散组比较,四慧合剂组EphB2的mRNA表达明显增多(P<0.01)。生慧汤组较模型组亦能明显增加NR2A的mRNA表达(P<0.01)。结论中枢疲劳能导致大鼠迷宫成绩下降和海马NR2B和EphB2基因表达下调。四慧合剂能增加大鼠的学习记忆能力,可能是通过上调大鼠海马NR2B和EphB2的mRNA表达所致。
Objective To study the changes of learning and memory and expression of NR2A, NR2B subunit and EphB2 receptor of hippocampal N-methyl D-aspartate (NMDA) receptor in rats with fatigue. San, Health Hui Tang, four-Hui mixture on its impact. Methods The multi-platform method was used to perform central sleep deprivation for 168 hours. The experimental animals were randomly divided into normal control group, model group, Sini Powder group, Shenghui Decoction group and Sihui Mixture group with 10 rats in each group. Sleep deprivation at 6,30,54,78,102,126,150 h time point intragastric administration and distilled water. The changes of learning and memory abilities in rats were observed by Y-maze. The mRNA expression of NR2A, NR2B subunits and EphB2 receptors in hippocampus of rats were quantitatively analyzed by real-time PCR. Results Compared with the normal control group, the correct percentage of Y maze in model group was significantly decreased (P <0.05), the mRNA expression of NR2B and EphB2 in hippocampus was significantly decreased (P <0.01), while the expression of NR2A did not change significantly. Compared with the model group, Sihui Mixture group can significantly improve the maze test results and mRNA expression of NR2A, EphB2 in the hippocampus (P <0.01), but there was no significant difference in the Sini Decoction group and Shenghui Decoction group. Compared with Sini Decoction, EphB2 mRNA expression in Sihui Mixture group increased significantly (P <0.01). Compared with the model group, Shenghui Decoction also significantly increased NR2A mRNA expression (P <0.01). Conclusion Central fatigue can lead to decreased maze results and down-regulated expression of NR2B and EphB2 genes in the hippocampus. Sihui Mixture can increase learning and memory in rats, probably by up-regulating the mRNA expression of NR2B and EphB2 in the hippocampus of rats.